January 31, 2008

Bathing Bolitophagus Beetles in Buffer: Brewing Bug Broth!

Today has been a busy and, apparently, alliterative day for me. The high point of my day so far is that I've finally started trying a non-destructive DNA extraction from dried beetles according to the protocol set forth in:

Gilbert MTP et al. (2007) DNA Extraction from Dry Museum Beetles without Conferring External Morphological Damage. PLoS ONE 2(3): e272. doi:10.1371/journal.pone.0000272

If all goes well, my dead Hungarian Bolitophagus reticulans specimens will have a very pleasant evening and, at about 11:00 AM tomorrow, I'll have a buffer solution containing DNA and still-whole specimens. I'm also running a control on two specimens from the same lot by using a standard, destructive protocol for extraction. I should be able to test not only whether Gilbert's protocol is non-destructive, but whether it's actually useful in 15 year old specimens. I've been trying to get to this all week!

I almost slipped up in concocting this devil's brew of a buffer. See, the stuff contains a chemical called dithiotreitol, which is used to break disulfide bonds in proteins. I've never worked with it before and, silly me, I forgot to check it's solubility which turns out to be quite low. Luckily, one of the professors here caught my error in time. As 5g of the stuff goes for well over $150, not wasting the stuff is quite a good idea. Put it this way: if my mistake wouldn't have been caught, I'd have used 100X more of the DTT than would have dissolved. Must remember to check the solubility of these things!

I really have been running like a madman this week. Even today started with my driving LL to the train station. She's back at FSU for her conference; I think she may even have landed by now. Besides spending most of the day in the lab, I also have to teach Intro Bio tonight until 9:00. Then I can maybe go to the supermarket and get some coffee and finally go home, eat and, if I'm awake enough, watch a bad horror flick to unwind (I've got something called Dead Clowns waiting; it's bound to be a classic).

Tomorrow I'll set up for amplifying bug DNA. Who knows... I could have it ready to sequence by early next week and become, as far as I know, the first person to ever bother sequencing the rDNA and a chitinase gene from B. reticulans. That is, if my chi-primers work. If not, I'll still have the rDNA, anyhow. Surely the nobel committee will be considering me next year for such an accomplishment.

Because, you know, everybody loves a fungus-eating beetle from Hungary, right? Ah, but it's only a first step on the long road to clarifying the phylogeny of the second-largest family under Coleoptera. That and a quarter will someday buy me a gumball.

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